Problem: Diabetes (DM) increases cardiovascular (CV) and all-cause mortality despite aggressive risk factor modification. Our long term goal is to define novel targets for cardiovascular risk reduction in diabetes. Vascular mitochondria are a potential new target. Mitochondria regulate endothelial function and smooth muscle cell (SMC) proliferation and mitochondrial dysfunction is a hallmark of diabetes. Exercise elicits an adaptive improvement in mitochondrial quality in healthy vessels; this response is absent in diabetes. The question addressed in this proposal is whether pharmacological targeting of eNOS and SIRT with glucagon- like peptide 1 (GLP-1) can restore signaling to mitochondrial biogenesis and improve mitochondrial dynamics and vascular function. GLP-1 is a diabetes medication that also induces eNOS, SIRT1, mitochondrial biogenesis and autophagy 21. New preliminary data suggest that saxagliptin (SAXA), a dipeptidyl peptidase-4 inhibitor that increases circulating endogenous GLP-1, restores induction of eNOS and mitochondrial protein expression with exercise in DM and improves running distance. Hypotheses: Abnormal mitochondrial function and impaired stress mediated mitochondrial dynamics in the diabetic vasculature will be improved by GLP-1 via eNOS and SIRT1 signaling. SA #1: What is the impact of GLP-1 on vascular mitochondrial adaptation in DM? Rationale: Control rats subjected to 8 day exercise show increased activation of the mitochondrial biogenesis, improved mitochondrial function plus increased fusion and decreased fission. We will examine the impact of GLP-1 on vascular mitochondrial function and turnover in diabetes and use targeted deletion of eNOS, endothelial cell (EC) SIRT1, SMC SIRT1 with or without exercise to test mitochondrial homeostatic adaptation in vivo. Hypothesis 1.1: Intervention with GLP-1 will improve mitochondrial function through augmentation of vascular mitochondrial adaptation (biogenesis, fusion and autophagy) to exercise in high fat induced diabetes. Hypothesis 1.2: eNOS and/or SIRT1 are required for GLP-1 rescue of mitochondrial adaptation. Approach: 1a. Male C57BL6 on chow versus high fat (HF) diet for 10 weeks will be randomized to GLP-1 or vehicle with or without exercise for 8 days. 1b. c57eNOS -/- and c57b eNOS +/+ will be treated as in SA1a. 1c. c57 mice with EC or SMC deletion of SIRT1 and controls will be treated as in SA1a. These experiments will clarify the importance of eNOS and SIRT1 for vascular mitochondrial adaptation to exercise in vivo and define the impact of diabetes and GLP-1 on these pathways. SA #2: How does diabetes affect dynamic mitochondrial adaptation in vascular cells in vitro? Rationale: Regulation of mitochondrial function requires a complex interplay between mitochondrial biogene- sis and remodeling through fission, fusion and autophagy. Hypothesis 2.1: Endothelial cells and/or smooth muscle cells from DM models will have impaired mitochondrial dynamics. Hypothesis 2.2: GLP-1 will improve mitochondrial function and dynamics in DM cells through eNOS and/or SIRT. Approach: These experiments will examine the impact of DM and GLP-1 on mitochondrial quality and dynamics in vitro and employ genetic approaches to define the relative contribution of eNOS and SIRT for mitochondrial quality in vascular cells. Impact on the Veteran Population: Mitochondrial dynamics are a novel target to decrease excess CV risk in diabetes. Interventions targeting mitochondrial ROS have been consistently ineffective in human studies. Our observation that 8 days of treatment with SAXA restores vascular induction of mitochondrial biogenesis provides proof of concept that impaired vascular mitochondrial adaptation is targetable with a currently available drug. Pharmacological restoration of mitochondrial dynamics in diabetes could have implications for macrovascular and microvascular complications of diabetes.